Detection and quantification of the toxic microalgae Karenia brevis using lab on a chip mRNA sequence-based amplification

Loukas, Christos-Moritz; McQuillan, Jonathan S.; Laouenan, Florian; Tsaloglou, Maria-Nefeli; Ruano-Lopez, Jesus M.; Mowlem, Matthew C.. 2017 Detection and quantification of the toxic microalgae Karenia brevis using lab on a chip mRNA sequence-based amplification. Journal of Microbiological Methods, 139. 189-195. 10.1016/j.mimet.2017.06.008

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Text © 2017 Elsevier B.V. This is the author’s version of a work that was accepted for publication in Journal of Microbiological Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was/will be published in Journal of Microbiological Methods (doi:10.1016/j.mimet.2017.06.008) Loukas et al 2017_REVISED.docx - Accepted Version Available under License Creative Commons Attribution Non-commercial No Derivatives 4.0. Download (7MB)

Abstract/Summary

Now and again, the rapid proliferation of certain species of phytoplankton can give rise to Harmful Algal Blooms, which pose a serious threat to marine life and human health. Current methods of monitoring phytoplankton are limited by poor specificity or by the requirement to return samples to a highly resourced, centralised lab. The Lab Card is a small, microfluidic cassette which, when used in tandem with a portable Lab Card Reader can be used to sensitively and specifically quantify harmful algae in the field, from nucleic acid extracts using RNA amplification; a sensitive and specific method for the enumeration of potentially any species based on their unique genetic signatures. This study reports the culmination of work to develop a Lab Card-based genetic assay to quantify the harmful algae Karenia brevis using mRNA amplification by the Nucleic Acid Sequence Based Amplification (NASBA) method. K. brevis cells were quantified by amplification of the rbcL gene transcript in nucleic acid extracts of K. brevis cell samples. A novel enzyme dehydration and preservation method was combined with a pre-existing reagent Gelification method to prepare fully preserved Lab Cards with a shelf-life of at least six weeks prior to use. Using an internal control (IC), the Lab Card-based rbcL NASBA was demonstrated for the quantification of K. brevis from cell extracts containing between 50 and 5000 cells. This is the first demonstration of quantitation of K. brevis using IC-NASBA on a Lab Card.

Item Type:	Publication - Article
Digital Object Identifier (DOI):	10.1016/j.mimet.2017.06.008
ISSN:	01677012
Additional Keywords:	Karenia brevis; NASBA; Internal control; Lab Card; Autonomous; Sensing
Date made live:	26 Jul 2017 08:40 +0 (UTC)
URI:	https://nora.nerc.ac.uk/id/eprint/517378

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